Chromatography membranes, devices containing them, and methods of use thereof

ABSTRACT

Described herein are fluid treatment devices for use in tangential flow filtration, comprising a housing unit and a composite material, wherein the composite material comprises: a support member comprising a plurality of pores extending through the support member; and a non-self-supporting macroporous cross-linked gel comprising macropores having an average size of 10 nm to 3000 nm, said macroporous gel being located in the pores of the support member. The invention also relates to a method of separating a substance from a fluid, comprising the step of placing the fluid in contact with an inventive device, thereby adsorbing or absorbing the substance to the composite material contained therein.

RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 12/551,762, filed Sep. 1, 2009, which claims the benefit of priorityto U.S. Provisional Patent Application Ser. No. 61/093,600, filed Sep.2, 2008; and U.S. Provisional Patent Application Ser. No. 61/102,797,filed Oct. 3, 2008.

BACKGROUND OF THE INVENTION

Membrane-based water treatment processes were first introduced in the1970s. Since then, membrane-based separation technologies have beenutilized in a number of other industries. In the pharmaceutical andbiotechnology industries, the use of preparative chromatography, directflow filtration (DFF) and tangential flow filtration (TFF), includingmicro-, ultra-, nano-filtration and diafiltration are well-establishedmethods for the separation of dissolved molecules or suspendedparticulates. Ultrafiltration (UF) and microfiltration (MF) membraneshave become essential to separation and purification in the manufactureof biomolecules. Biomolecular manufacturing, regardless of its scale,generally employs one or more steps using filtration. The attractivenessof these membrane separations rests on several features including, forexample, high separation power, and simplicity, requiring only theapplication of pressure differentials between the feed stream and thepermeate. This simple and reliable one-stage filtering of the sampleinto two fractions makes membrane separation a valuable approach toseparation and purification.

Notably, the separation and recovery of biomolecules, such as enzymesand glycoproteins, are critical cost-determining steps in most of thedown-stream processes in the biotechnology industry. For example,separation of lysozyme from crude sources, such as egg white, has beenachieved by salt precipitation (U.S. Pat. No. 4,504,583), or ionexchange techniques (U.S. Pat. Nos. 4,705,755; 4,966,851; 4,518,695; and4,104,125). Due to the viscous, highly concentrated nature of egg white,and the nature of the other protein constituents, recovering high-puritylysozyme in good yield is extremely laborious and costly.

In one class of membrane separations, the species of interest is thatwhich is retained by the membrane, in which case the objective of theseparation is typically to remove smaller contaminants, to concentratethe solution, or to affect a buffer exchange using diafiltration. Inanother class of membrane separations, the species of interest is thatwhich permeates through the filter, and the objective is typically toremove larger contaminants. In MF, the retained species are generallyparticulates, organelles, bacteria or other microorganisms, while thosethat permeate are proteins, colloids, peptides, small molecules andions. In UF the retained species are typically proteins and, in general,macromolecules, while those that permeate are peptides, ions and, ingeneral, small molecules.

In “dead-end,” “normal flow,” or “direct flow” filtration (DFF), afiltration device is used that has one inlet and one outlet. The total(100%) solution volume is forced through a porous filter. DFF devicesare commonly single-use devices. Such membrane filters or depth filtersare commercially available in different filter area sizes as well asdifferent pore sizes. Depending upon the selected pore size, moleculesor particulates smaller than the average membrane pore size will pass(together with solvent) through the filter. Thus, direct flow filtration(DFF) devices allow for the selective removal of particulates, bacteria,viruses, cell debris, and large macromolecules.

Conventional filters in which all of the fluid entering the filterhousing passes through the filter element (DFF) typically operate at lowshear near the surface of the filter medium. Thus, when a highlyflocculating dispersion is delivered into a conventional filter deviceby a conventional delivery system, flocs ordinarily tend to form nearthe surface of the filter medium. The flow field moves the flocs ontothe surface and into the bulk of the filter medium, ultimately resultingin plugging of the filter. In practice, a plugged filter may cause asignificant amount of downtime for a filter change.

A raw or semi-conditioned process stream that contains high-valuematerials is often highly viscous or highly contaminated. As such, DFFseparation approaches are difficult or challenging due to blinding ofthe membrane with the solute present in the feed stream. Additionally,these processes often require high pressure to maintain a reasonableflux of permeate.

In contrast, tangential flow filtration (TFF) devices, also known ascross-flow filtration devices, have one inlet, one retentate outlet andat least one permeate outlet. Tangential flow denotes a filtrationconfiguration in which a flowing fluid is directed along the surface ofa filter medium, substantially parallel (tangential) to the surface ofthe filter medium. In this configuration, the solute adsorbs or absorbsto the surface or the pores of the membrane as the eluent flows over thesurface. The purified portion of fluid that passes through such filtermedium has a velocity component which is “cross-wise”, i.e.,perpendicular to the direction of the fluid flowing along the surface ofsuch filter medium. In TFF, the retentate (or decantate) can berepeatedly re-circulated with the objective of improving filtrationefficiency and enhancing the permeate yield. The re-circulated retentatesolution pathway runs parallel to the membrane surface and is pumpedpast the membrane with sufficient velocity to ensure a surface cleaningaction. However, only a relatively small amount of permeate is collectedduring each retentate volume-pass, and thus a significant processingtime is typically associated with TFF procedures. If an appropriatemembrane is selected for a specific separation, a second liquid can beused to elute the material adsorbed or absorbed to the membrane forharvesting.

Crossflow filtration or tangential filtration is a well known filtrationprocess. Reference may be had e.g., to U.S. Pat. Nos. 5,681,464,6,461,513; 6,331,253, 6,475,071, 5,783,085, 4,790,942, the disclosuresof which are incorporated herein by reference. Reference may also be hadto “Filter and Filtration Handbook”, 4th Ed., T. Christopher Dickenson,Elsevier Advanced Technology, 1997, the disclosure of which isincorporated herein by reference.

In TFF careful attention must be paid in the device design, as flowdynamics play an important role in the efficiency of the system.Turbulent flow must be minimized in these systems, so as to notphysically disassociate a desired substance from the membrane surface.Turbulence is flow dominated by recirculation, eddies, and apparentrandomness. Flow in which turbulence is not exhibited is called laminar.A steady, laminar flow is desired.

For optimal results, both DFF and TFF demand careful attention to filterporosity and filter area, as well as required differential pressures andselected pump rates. However, filtration devices tend to clog when usedover an extended period of time and must be timely replaced. Clogging ofa filtration device occurs: (1) when the membrane pores becomeobstructed, typically with trapped cells, particulate matter, celldebris or the like, or (2) when the feed channel (into a TFF device)becomes obstructed by solids or colloidal material and/or cell debris.This clogging of the feed channel or membrane pores results in adecreased liquid flow across the porous filter membrane. The result is achange in system pressure which, if not properly addressed, runs therisk of serious detriment to the operation which incorporates thefiltration procedure.

As such, the choice of membrane in each of the filtration techniques iscritical to the efficiency and success of the separation. Compositemembranes with high specificity and high binding capacity have beendescribed in U.S. Pat. No. 7,316,919, and US Patent ApplicationPublication Nos. 2008/0314831 and 2008/0312416, which are herebyincorporated by reference in their entirety. These materials are highlyversatile and can be designed for specific separation situations.

A wide variety of devices are available for these applications.Typically, devices are categorized by configuration into categoriesincluding the following: flat plate (for example, cassette or plate andframe), spiral (or spiral wound), tubular, or hollow fiber. The choiceof device configuration is driven by reliability, performance, and costfor each specific application.

Flat plate or cassette devices consist of membranes cast on plates; theplates are then reliably stacked. The devices may or may not haveflexible screens in the feed channels to support the membranes. Anappealing advantage of a configuration such as this is its very compactdesign. However, channel height control, defined by plate-to-plateinteraction and distance, must be very carefully considered

Tubular devices consist of a membrane cast on the inside surface oroutside diameter of a porous support tube. Typically, a feed solution ispumped through the center of the tube at velocities as high as 20 ft/s.These cross-flow velocities minimize the formation of a concentrationpolarization layer on the membrane surface, promoting high and stableflux and easy cleaning. The permeate is driven through the membrane.Despite the apparent advantages of using a system such as this, the costtends to be high.

Spiral-wound devices consist of multiple layers of folded membrane, feedscreen, and permeate screen wound around a center permeate collectiontube (FIG. 23). Typically found in water purification applications,these devices are also compact and can operate at low pressure to saveenergy, but are suitable for high pressure applications as well. Thecost per membrane area is typically low.

Typical spiral wound filters consist of about 1 to about 6 spiral woundelements coupled in a serial flow mode and placed in a cylindricalpressure vessel. Between two membranes in the roll is placed a permeableporous medium for conduction of fluid, the concentrate spacer, to ensurethat the concentrate can flow over the membrane in order to bedistributed all over the surface and to continuously rinse the membranefrom accumulating solids. The filter elements are kept tightly wound bya hard, impermeable shell. In this configuration flow in and out of thefilter element will be through the ends in an axial direction.

An unmet need exists in many applications where high contaminate feedstreams will immediately plug or blind the membrane media in a typicalDFF mode or, when the membranes employed are incapable of anyappreciable substrate capture, in cross-flow modes. Utilizing versatile,high performance, high throughput membranes capable of high bindingcapacities in filtration devices would provide separation systems withperformances far exceeding any known technology in a variety of artareas.

SUMMARY OF THE INVENTION

In certain embodiments, the invention relates to a fluid treatmentdevice comprising

-   -   a housing unit, wherein the housing unit comprises    -   (a) an inlet and an outlet;    -   (b) a fluid flow path between the inlet and the outlet; and    -   (c) a composite material within the housing unit, wherein the        composite material comprises        -   a support member comprising a plurality of pores extending            through the support member; and        -   a non-self-supporting macroporous cross-linked gel            comprising macropores having an average size of 10 nm to            3000 nm, said macroporous gel being located in the pores of            the support member;        -   wherein said macropores of said macroporous cross-linked gel            are smaller than said pores of said support member;            wherein the pores of the support member are substantially            perpendicular to the fluid flow path.

In certain embodiments, the invention relates to a fluid treatmentdevice comprising

-   -   a plurality of housing units, wherein each housing unit        comprises    -   (a) an inlet and an outlet;    -   (b) a fluid flow path between the inlet and the outlet; and    -   (c) a composite material within the housing unit, wherein the        composite material comprises        -   a support member comprising a plurality of pores extending            through the support member; and        -   a non-self-supporting macroporous cross-linked gel            comprising macropores having an average size of 10 nm to            3000 nm, said macroporous gel being located in the pores of            the support member;        -   wherein said macropores of said macroporous cross-linked gel            are smaller than said pores of said support member; and            wherein the pores of the support member are substantially            perpendicular to the fluid flow path.

In certain embodiments, the invention relates to any one of theaforementioned fluid treatment devices, wherein the composite materialis arranged in a substantially coplanar stack of substantiallycoextensive sheets, a substantially tubular configuration, or asubstantially spiral wound configuration.

In certain embodiments, the invention relates to any one of theaforementioned fluid treatment devices, wherein the macroporouscross-linked gel is a neutral or charged hydrogel, a polyelectrolytegel, a hydrophobic gel, a neutral gel, or a gel comprising functionalgroups.

In certain embodiments, the invention relates to any one of theaforementioned fluid treatment devices, wherein said functional groupsare selected from the group consisting of amino acid ligands, antigenand antibody ligands, dye ligands, biological molecules, biologicalions, and metal affinity ligands.

In certain embodiments, the invention relates to any one of theaforementioned fluid treatment devices, wherein said functional groupsare metal affinity ligands. In certain embodiments, the inventionrelates to any one of the aforementioned fluid treatment devices,further comprising a plurality of metal ions complexed to a plurality ofsaid metal affinity ligands. In certain embodiments, the inventionrelates to any one of the aforementioned fluid treatment devices,wherein said metal affinity ligands are iminodicarboxylic acid ligands;and said metal ions are nickel.

In certain embodiments, the invention relates to any one of theaforementioned fluid treatment devices, wherein said functional groupsare biological molecules or biological ions. In certain embodiments, theinvention relates to any one of the aforementioned fluid treatmentdevices, wherein said functional groups are Protein A.

In certain embodiments, the invention relates to a method comprising thestep of:

-   -   contacting a first fluid comprising a substance with a composite        material in any one of the aforementioned fluid treatment        devices, thereby adsorbing or absorbing the substance onto the        composite material.

In certain embodiments, the invention relates to any one of theaforementioned methods, further comprising the step of

-   -   placing the first fluid in an inlet of the fluid treatment        device.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the first fluid is passed along a fluidflow path substantially perpendicular to the pores of the supportmember.

In certain embodiments, the invention relates to any one of theaforementioned methods, further comprising the step of

-   -   contacting a second fluid with the substance adsorbed or        absorbed onto the composite material, thereby releasing the        substance from the composite material.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the first fluid is a suspension of cellsor a suspension of aggregates.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the substance is a biological moleculeor biological ion. In certain embodiments, the invention relates to anyone of the aforementioned methods, wherein the biological molecule orbiological ion is a protein; and the protein comprises exposed His aminoacid residues. In certain embodiments, the invention relates to any oneof the aforementioned methods, wherein the biological molecule orbiological ion is a monoclonal antibody.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the substance is a metal-containingparticle, or a metal-containing ion.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the first fluid is waste water.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the first fluid comprises egg white; andthe substance is lysozyme.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts the results of dead-end compared to cross-flow modes forviral capture using an ion-exchange membrane. In both cases, the lowervalue obtained with dead-end flow was due to fouling of the membraneduring the experiment.

FIG. 2 depicts a chromatogram from the elution of a mixture proteins(ovalbumin and lysozyme) captured directly from unprocessed egg whitesusing ion-exchange membranes in cross-flow mode. The results demonstratethat proteins can be selectively removed from an unprocessed, highlyviscous feed stream.

FIG. 3 depicts chromatograms of eluent fluids from egg white-loadedion-exchange membranes. Curves show selectivity of elution, based onbuffer (saline solution) selection. This series of curves demonstratesthe ability selectively to separate (i.e., chromatographically) capturedtarget materials in high purity, or as mixtures essentially free fromother constituents.

FIG. 4 depicts a schematic of the cross-flow and capture steps (top andmiddle), and a trans-membrane collection step (bottom).

FIG. 5 depicts the effects of wrap design on device performance showingthat coarse mesh spacers lead to improved performance in separatinglysozyme from egg white using ion-exchange membrane.

FIG. 6 depicts a wrapped column device inserted into a housing. Inletcap not shown for clarity.

FIG. 7 depicts a simplified cross-section of a cassette showingtrans-membrane flow of target material as indicated by arrows.Harvesting could also be accomplished by cross-flow, if required, usinga fluid that selectively eluted the bound target materials.

FIG. 8 depicts a schematic illustration of a typical cassette design.Flow is shown with trans-membrane harvesting that is occurringsimultaneously with a flowing feed stream.

FIG. 9 depicts a cross-section of a disposable or semi-disposablehousing for a 25 mm syringe column comprising a disk-shaped filtrationmembrane for lab-scale use.

FIG. 10 depicts top view (top left), side view (right), and actual sizeviews (bottom left) of the outlet half of the syringe tip filters foruse in the housing shown in FIG. 9 and FIG. 11. All units in thedrawings are in inches.

FIG. 11 depicts a cross-section of the disposable or semi-disposablehousing for a 25 mm syringe column comprising a disk-shaped filtrationmembrane for lab-scale use.

FIG. 12 depicts a cross-section of the disposable or semi-disposablehousing for a 50 mm syringe column comprising a disk-shaped filtrationmembrane for lab-scale use.

FIG. 13 depicts the drainage grid in the housing depicted in FIG. 12 foruse with a 50 mm disk-shaped membrane.

FIG. 14 depicts an inlet flow deflector in a reusable stainless steelhousing for a 50 mm syringe column comprising a disk-shaped filtrationmembrane for lab-scale use.

FIG. 15 depicts a stainless steel holder for use as a reusable housingfor a 25 mm disk-shaped membrane for lab-scale use.

FIG. 16 depicts a component of a reusable stainless steel housing forlab-scale syringe columns.

FIG. 17 depicts two components of a reusable stainless steel housing forlab-scale syringe columns.

FIG. 18 depicts a maxi spin column (left), and a device for supporting acut disk membrane within the column (right).

FIG. 19 depicts the dimensions of a maxi spin column.

FIG. 20 depicts the dimensions of a maxi spin column with a device forsupporting a cut disk membrane within the column.

FIG. 21 depicts a mini spin column with a device for supporting a cutdisk membrane within the column.

FIG. 22 depicts the dimensions of a mini spin column with a device forsupporting a cut disk membrane within the column.

FIG. 23 depicts an exemplary configuration of a spiral wound device.There are three series of concentric envelopes, wherein each envelopehas a spacer material inside and three of the sides are sealed. Eachenvelope is separated by a feed spacer. Fluid flow is directed such thatraw fluid travels on the outside of each envelope and is forced throughthe membrane. The permeate travels along the permeate spacer to thepermeate collection pipe.

FIG. 24 depicts an exemplary synthetic scheme for incorporation of ametal affinity ligand into the membrane. In this case, the metalaffinity ligand is the sodium salt of iminodiacetic acid (IDA(Na)₂).

FIG. 25 depicts an exemplary synthetic scheme for incorporation of ametal affinity ligand into the membrane. In this case, the metalaffinity ligand is ethylenediamine (EDA).

FIG. 26 depicts an exemplary synthetic scheme for incorporation of ametal affinity ligand into the membrane. In this case, the metalaffinity ligand is hexamethylenediamine (HMDA).

FIG. 27 depicts an exemplary synthetic scheme for incorporation of ametal affinity ligand into the membrane. In this case, the metalaffinity ligand is diethanolamine.

FIG. 28 depicts an exemplary synthetic scheme for incorporation of ametal affinity ligand into the membrane. In this case, the metalaffinity ligand is pentaethylenehexamine (PEHA).

FIG. 29 depicts an exemplary synthetic scheme for incorporation of ametal affinity ligand into the membrane. In this case, the metalaffinity ligand is triethylenetetramine (TETA).

FIG. 30 depicts an exemplary synthetic scheme for incorporation of ametal affinity ligand into the membrane. In this case, the metalaffinity ligand is the sodium salt of tris(carboxymethyl)ethylenediamine (TED(Na)₃).

DETAILED DESCRIPTION OF THE INVENTION

Overview

Disclosed is a hydrophilic, high binding, high throughput chromatographymembrane that is effective for selective capture of target materials,such as bio-molecules, from raw or dirty process streams. This captureprocess can be accomplished by binding of the target molecules at thesurface or near surface of the membrane media (“cross-flow” mode), asopposed to the more typical trans-membrane mode. The captured targetspecies can be collected in a highly purified form in subsequentprocedures. These final steps are chromatographic in nature and allowfor controlled separation of the target materials. Importantly, thecollection step and the separation step can be done in either tangentialflow or in trans-membrane flow or combinations thereof. See, e.g., FIG.1.

Exemplary device designs suitable for this process include those inwhich the membrane is incorporated into a modified cassette, wrap, orspiral-wound cross-flow separation device designed for low-shearfluid-flow, and minimization of uncontrolled or undesired trans-membraneflow. For example, such devices were found to be effective forseparating proteins or viruses from highly viscous and or highlycontaminated feed streams with a minimum of process fluid flux acrossthe membrane. The cross flow (tangential flow) format allows for greaterflexibility in washing and eluting the target molecule(s). The crossflow devices can be run in feed-to-retentate mode and perform a surfaceion exchange or affinity separation. Washing can be done infeed-to-retentate mode, feed-to-permeate or permeate-to-feed mode, or ina sequential mode.

The incorporation of the hydrophilic, high performance chromatographymembrane into a modified cross flow device provides a separation devicethat purifies target molecules from highly viscous or high particulatefeed streams, and completes both clarification and capture of targetspecies with no intervening steps. Moreover, the materials andconstructs described here do not preclude the use of the same membranematerials in traditional device designs, such as pleated dead-endcapsules. Importantly, these products can produce highly purifiedproteins, vaccines, or nutraceuticals from feed streams that cannot beprocessed directly with current commercial technology. Additionally, thedevices and methods of the present invention allow for faster processingof large volumes of feed streams than any current technology.

For example, due to the viscous, highly concentrated nature of eggwhite, typical filtration schemes prove to be problematic when trying tocollect constituents present in relatively low concentrations. Using thedevices and methods of the present invention, lysozyme can be easilyseparated from egg white with high recovery and high purity.

In certain embodiments, the invention relates to a device that displayssuperior performance in comparison to know devices. In certainembodiments, the devices may tolerate about 10× to about 100× higherthroughput than resins. In certain embodiments, the devices may displayup to about 25× higher binding capacity than existing chromatographicmembranes and resins.

In certain embodiments, the invention relates to a device that isscalable and produces predictable results in the transitions from Lab toPilot to Production, unlike conventional resin products.

In certain embodiments, the invention relates to a device thatencompasses a robust technology. In certain embodiments, the superiormechanical strength of the devices and the inherent hydrophilicity ofthe composite membranes lead to longer in-process product lifetimes andmore consistent performance.

In certain embodiments, the invention relates to a device that may beavailable as a single use or multi-cycle disposable unit. Thisflexibility may eliminate costly and time consuming cleaning and storagevalidation. Furthermore, the devices of the invention enable simpleprocess and may improve regulatory compliance.

In certain embodiments, the invention relates to separation processesthat may require reduced buffer usage. In certain embodiments, usingdevices of the present invention may eliminate the need for columncleaning, equilibration, or storage in expensive buffers. In certainembodiments, the devices of the invention may tolerate higherconcentration feed stream, so no dilution may be needed.

In certain embodiments, using the devices described herein may lowercapital expenses and may offer significant operational cost savings fora client. In certain embodiments, the devices of the invention may havea lower initial cost and faster delivery. In certain embodiments, thedevices allow for lower staffing requirements and reduced maintenancecosts.

In certain embodiments, the invention relates to a device with a smallfootprint. In certain embodiments, the devices of the invention exhibithigher binding capacity and require less floor space than typical resinbed chromatography devices.

Definitions

For convenience, before further description of the present invention,certain terms employed in the specification, examples and appendedclaims are collected here. These definitions should be read in light ofthe remainder of the disclosure and understood as by a person of skillin the art. Unless defined otherwise, all technical and scientific termsused herein have the same meaning as commonly understood by a person ofordinary skill in the art.

In describing the present invention, a variety of terms are used in thedescription. Standard terminology is widely used in filtration, fluiddelivery, and general fluid processing art.

The articles “a” and “an” are used herein to refer to one or to morethan one (i.e., to at least one) of the grammatical object of thearticle. By way of example, “an element” means one element or more thanone element.

The term “associated with” as used herein in such phrases as, forexample, “an inorganic metal oxide associated with an stabilizingcompound,” refers to the presence of either weak or strong or bothinteractions between molecules. For example weak interactions mayinclude, for example, electrostatic, van der Waals, or hydrogen-bondinginteractions. Stronger interactions, also referred to as beingchemically bonded, refer to, for example, covalent, ionic, orcoordinative bonds between two molecules. The term “associated with”also refers to a compound that may be physically intertwined within thefoldings of another molecule, even when none of the above types of bondsare present. For example, an inorganic compound may be considered asbeing in association with a polymer by virtue of it existing within theinterstices of the polymer.

The terms “comprise” and “comprising” are used in the inclusive, opensense, meaning that additional elements may be included.

The term “including” is used to mean “including but not limited to.”“Including” and “including but not limited to” are used interchangeably.

The term “polymer” is used to mean a large molecule formed by the unionof repeating units (monomers). The term polymer also encompassescopolymers.

The term “co-polymer” is used to mean a polymer of at least two or moredifferent monomers. A co-polymer can be comprised of a cross-linker anda monomer, if the cross-linker is a difunctional monomer.

The term “two phase fluid” is used to mean a fluid comprising a liquidphase in which either substantially solid particles are dispersedtherethrough, or a first liquid phase in which droplets or particles ofa second liquid phase immiscible with such first liquid phase aredispersed through such first liquid phase. A “multiphase fluid” is usedto mean a fluid comprising a first liquid phase in which at least oneadditional second solid or liquid phase is dispersed therethrough.

The term “particle” is used to mean a discreet liquid droplet or a solidobject, with a characteristic dimension such as a diameter or length ofbetween about one nanometer, and about one-tenth of a meter.

The term “particle size” is used to mean a number-average orweight-average particle size as measured by conventional particle sizemeasuring techniques well known to those skilled in the art, such asdynamic or static light-scattering, sedimentation field-flowfractionation, photon-correlation spectroscopy, or disk centrifugation.By “an effective average particle size of less than about 1000 nm” it ismeant that at least about 90% of the particles have a number-average orweight-average particle size of less than about 1000 nm when measured byat least one of the above-noted techniques. The particular size ofparticles in a fluid being processed will depend upon the particularapplication.

The term “interstices” is used to mean a space, especially a small ornarrow one, between things or parts.

The term “dispersion” is used to mean any fluid comprising a liquidphase in which substantially solid particles are suspended, and remainsuspended, at least temporarily.

The term “slurry” is used to mean any fluid comprising a liquid phase inwhich substantially solid particles are present. Such particles may ormay not be suspended in such fluid.

The term “emulsion” is used to mean any fluid comprising a first liquidphase within which droplets or particles of a substantially liquidsecond phase are suspended, and remain suspended, at least temporarily.In reference to discreet entities of a second liquid phase in a firstliquid phase, the terms “droplets” and “particles” are usedinterchangeably herein.

The term “crossflow” in reference to filtration is used to mean afiltration configuration in which a flowing fluid is directed along thesurface of a filter medium, and the portion of fluid that passes throughsuch filter medium has a velocity component which is “cross-wise”, i.e.,perpendicular to the direction of the fluid flowing along the surface ofsuch filter medium.

The term “tangential filtration” is used to mean a filtration process inwhich a flowing fluid is directed substantially parallel (i.e.,tangential) to the surface of a filter medium, and a portion of fluidpasses through such filter medium to provide a permeate. The terms“tangential filtration” and “crossflow filtration” are often usedinterchangeably in the art.

The term “permeate” is used to mean the portion of the fluid that passesthrough the filter medium and out through a first outlet port in thefilter device that is operatively connected to such filter medium. Theterm “decantate” is used to mean the portion of the fluid that flowsalong the surface of the filter medium, but does not pass through suchfilter medium, and passes out through a second outlet port in the filterdevice that is operatively connected to such filter medium.

Crossflow filtration and tangential filtration are well known filtrationprocesses. Reference may be had to, e.g., U.S. Pat. Nos. 5,681,464,6,461,513; 6,331,253, 6,475,071, 5,783,085, 4,790,942, the disclosuresof which are incorporated herein by reference. Reference may also be hadto “Filter and Filtration Handbook”, 4th Ed., T. Christopher Dickenson,Elsevier Advanced Technology, 1997, the disclosure of which isincorporated herein by reference.

The term “egg white” refers to the clear, aqueous liquid containedwithin an egg, as opposed to the yellow egg yolk. Egg white typicallycomprises about 15% proteins dissolved or suspended in water. Egg whiteproteins typically include ovalbumin, ovotransferrin, ovomucoid,globulins, lysozyme, ovomucin, and avidin.

Exemplary Devices

General Device Properties

In certain embodiment, the invention relates to a fluid treatment devicecomprising

-   -   a housing unit, wherein the housing unit comprises    -   (a) an inlet and an outlet;    -   (b) a fluid flow path between the inlet and the outlet; and    -   (c) a composite material within the housing unit, wherein the        composite material comprises        -   a support member comprising a plurality of pores extending            through the support member; and        -   a non-self-supporting macroporous cross-linked gel            comprising macropores having an average size of 10 nm to            3000 nm, said macroporous gel being located in the pores of            the support member;        -   wherein said macropores of said macroporous cross-linked gel            are smaller than said pores of said support member; and            wherein the pores of the support member are substantially            perpendicular to the fluid flow path.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the composite materialis arranged in a substantially coplanar stack of substantiallycoextensive sheets, a substantially tubular configuration, or asubstantially spiral wound configuration.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the composite materialhas 2 to 10 separate support members.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein

-   -   the support member is in the form of hollow porous fibers;    -   each hollow porous fiber defines a lumen;    -   the lumen is from about 20 μm to about 100 μm in diameter; and    -   the lumen is substantially perpendicular to the pores in the        hollow porous fiber support member.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein a plurality of hollowporous fibers are arranged in a bundle.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the bundle is encasedin a shell or a vessel.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein a plurality of bundlesis encased in a shell or a vessel.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the hollow porous fibercomprises a cap, a plug, or a seal. In certain embodiments, theinvention relates to any one of the above-mentioned fluid treatmentdevices, wherein the hollow porous fiber comprises a cap, a plug, or aseal on both ends. In certain embodiments, the end of the hollow porousfiber is “potted” such that the inside of the fiber is isolated from theoutside of the fiber. In certain embodiments, this is accomplishedthrough the use of a tubesheet. In certain embodiments, the pottingmaterial to form the tubesheet may be comprised of any suitablematerial. In certain embodiments, the potting material can be in liquidform when preparing the tubesheet and can thereafter be solidified,e.g., by cooling, curing, or the like. In certain embodiments, thesolidified potting material should exhibit sufficient structuralstrength for providing a tubesheet and be relatively inert moieties towhich it will be exposed during fluid separation operation. In certainembodiments, the potting material may be organic material (for example,epoxy), inorganic material, or organic material containing inorganicmaterial, and the potting material may be natural or synthetic. Incertain embodiments, typical inorganic materials include glasses,ceramics, cermets, metals and the like.

In certain embodiments, the hollow porous fiber may be of any convenientconfiguration. In certain embodiments, the hollow porous fiber iscircular, hexagonal, trilobal, or the like in cross-section and may haveridges, grooves, or the like extending inwardly or outwardly from thewalls of the hollow porous fibers. In certain embodiments, the hollowporous fiber may have an inner diameter of about 20 microns to about 200microns. In certain embodiments, the hollow porous fiber may have aninner diameter of about 40 microns. In certain embodiments, the hollowporous fiber may have a hollow ratio (being the area of the fiber boredivided by the area of the total cross-section of the fiber) of about10% to about 50% percent. In certain embodiments, the hollow porousfiber may have a hollow ratio of about 20%. In certain embodiments, thehollow porous fiber may be fabricated from various polymers such ascellulose, cellulose esters, cellulose ethers, polyamides, siliconeresins, polyurethane resins, unsaturated polyester resins or the like,or ceramics.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the composite materialis a pleated membrane.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit issubstantially cylindrical. In certain embodiments, the housing unit hasan inner diameter of from about 5 cm to about 50 cm.

In certain embodiments, the thickness of the walls of the housing unitmay be adapted to the specific operation conditions.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit isdisposable or reusable.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit isplastic or stainless steel.

In certain embodiments, the invention relates to a fluid treatmentdevice comprising a housing unit, wherein the housing unit comprises

at least one inlet and at least one outlet; and

a fluid flow path between the inlet and the outlet; wherein any one ofthe above-mentioned fluid treatment elements is across the fluid flowpath.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the composite materialcomprises:

-   -   (a) a support member comprising a plurality of pores extending        through the support member; and    -   (b) a non-self-supporting macroporous cross-linked gel        comprising macropores having an average size of 10 nm to 3000        nm, said macroporous gel being located in the pores of the        support member;    -   wherein        -   said macroporous cross-linked gel is present in the pores of            the support member in an amount sufficient such that, in            use, liquid passing through the composite material passes            through said macropores of said macroporous cross-linked            gel;        -   said macropores of said macroporous cross-linked gel are            smaller than said pores of said support member;        -   the support member is in the form of hollow porous fibers;        -   each hollow porous fiber defines a lumen;        -   the lumen is from about 20 μm to about 100 μm in diameter;            and        -   the lumen is substantially perpendicular to the pores in the            hollow porous fiber support member.

In certain embodiments, the fluid treatment devices comprise theabove-mentioned composite material, wherein a plurality of hollow porousfibers is arranged in a bundle. In certain embodiments, the fluidtreatment devices comprise the above-mentioned composite material,wherein the bundle is encased in a shell. In certain embodiments, thefluid treatment devices comprise the above-mentioned composite material,wherein the bundle is encased in a vessel. In certain embodiments, thefluid treatment devices comprise the above-mentioned composite material,wherein a plurality of bundles is encased in a shell. In certainembodiments, the fluid treatment devices comprise the above-mentionedcomposite material, wherein a plurality of bundles is encased in avessel.

In certain embodiments, the invention relates to a fluid treatmentdevice comprising

-   -   a plurality of housing units, wherein each housing unit        comprises    -   (a) an inlet and an outlet;    -   (b) a fluid flow path between the inlet and the outlet; and    -   (c) a composite material within the housing unit, wherein the        composite material comprises        -   a support member comprising a plurality of pores extending            through the support member; and        -   a non-self-supporting macroporous cross-linked gel            comprising macropores having an average size of 10 nm to            3000 nm, said macroporous gel being located in the pores of            the support member;        -   wherein said macropores of said macroporous cross-linked gel            are smaller than said pores of said support member; and            wherein the pores of the support member are substantially            perpendicular to the fluid flow path.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the plurality ofhousing units are arranged in series.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, comprising from about 2 toabout 10 housing units.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the composite materialis arranged in a substantially coextensive stack of substantiallycoplanar sheets; a substantially tubular configuration; or asubstantially spiral wound configuration.

In certain embodiments, wherein the inlet or the outlet is a press fitattachment point, a luer lock attachment point, or a hose barbattachment point. In certain embodiments, the inlet is a press fit, luerlock, or hose barb attachment points. In certain embodiments, the outletis a press fit, luer lock, or hose barb attachment points. In certainembodiments, the inlet and the outlet are different kinds of attachmentpoints from one another. In certain embodiments, the inlet and theoutlet are both press fit attachment points. In certain embodiments, theinlet and the outlet are both luer lock attachment points. In certainembodiments, the inlet and the outlet are both hose barb attachmentpoints.

Laboratory-Scale

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the composite materialis a cut disk membrane. In certain embodiments, the cut disks areintended to be used in re-usable housings. In certain embodiments, thecut disks are intended to be used in disposable housings. In certainembodiments, the cut disk membrane is substantially circular in shape.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the composite materialis a cut disk membrane. In certain embodiments, the cut disk membrane isfrom about 15 to about 60 mm in diameter. In certain embodiments, thecut disk membrane is from about 20 to about 55 mm in diameter. Incertain embodiments, the cut disk membrane is about 25 mm in diameter.In certain embodiments, the cut disk membrane is about 50 mm indiameter. For visualization of certain embodiments, see FIGS. 9-17.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit is asyringe tip. The term “syringe column” is used interchangeably with theterm “syringe tip.”

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit is asyringe column; and the composite material is in the form of a cut disk.In certain embodiments, the syringe column housing unit issemi-disposable.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit is aspin column. In certain embodiments, the invention relates to any one ofthe above-mentioned fluid treatment devices, wherein the housing unit isa spin column; and the composite material is in the form of a cut disk.A spin column is a tube with an upper and a lower half. The lower halfis closed at the bottom. In between the two halves is a cut diskmembrane held or suspended in some manner. A user loads the top halfwith a liquid containing the target (or contaminate) solute and placesthe spin column into a centrifuge. The centrifuge forces the liquidthrough the membrane when run at sufficient RPM. Once removed from thecentrifuge, the lower half of the device can be removed and the liquidcollected (if needed) or the top half can be eluted with additionalbuffer to remove the retained solute. In certain embodiments, the spincolumns can be made in many sizes.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit is aspin column; and the spin column has a capacity of from about 0.1 mL toabout 60 mL. In certain embodiments, the volume of the spin column referto the quantity of feed stream that may be processed by an exemplaryfluid treatment device. In certain embodiments, the spin column has acapacity of from about 0.3 mL to about 55 mL. In certain embodiments,the spin column has a capacity of about 0.5 mL. In certain embodiments,the spin column has a capacity of about 2 mL. In certain embodiments,the spin column has a capacity of up to about 50 mL. For visualizationof certain embodiments, see, e.g., FIGS. 18-22.

Process- and Manufacturing-Scale

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit is acassette configuration.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit is atubular configuration.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit is aspiral wound configuration.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unit is aplate and frame configuration.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices, wherein the housing unitcomprises a fluid treatment element, wherein the fluid treatment elementcomprises a hollow porous membrane.

Exemplary Fluid Treatment Elements

In certain embodiments, the invention relates to fluid treatmentelements. In certain embodiments, the fluid treatment element is acartridge for use in a fluid treatment device of the present invention.In certain embodiments, the invention relates to fluid treatmentelements comprising membranes. In certain embodiments, the inventionrelates to fluid treatment elements comprising composite materials foruse as membranes.

In certain embodiments, the fluid treatment elements are disposable orreusable.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment elements, wherein the element comprisesa hollow, generally cylindrical form.

In certain embodiments, the fluid treatment elements of the presentinvention accommodate high solid density materials. In certainembodiments, the fluid treatment elements of the present invention areused for their strength. In certain embodiments, the fluid treatmentelements of the present invention are used in heavy duty applications.In certain embodiments, the fluid treatment elements of the presentinvention can tolerate elevated temperatures for sustained periods.

In certain embodiments, the fluid treatment elements of the presentinvention exhibit reduced capture time in chromatography applications.In certain embodiments, the fluid treatment elements of the presentinvention exhibit high binding capacities.

Exemplary Composite Materials

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements comprising acomposite material. In certain embodiments, the invention comprises acomposite material for use as a membrane.

In certain embodiments, the composite materials used as membranes in thepresent invention are described in U.S. Pat. No. 7,316,919; and U.S.patent application Ser. Nos. 11/950,562, 12/108,178, 12/244,940,12/250,861, 12/211,618, and 12/250,869; all of which are herebyincorporated by reference.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein themacroporous crosslinked gel of the composite material has macropores ofaverage size between about 25 nm and about 1500 nm. In certainembodiments, the macroporous crosslinked gel has macropores of averagesize between about 50 nm and about 1000 nm. In certain embodiments, themacroporous crosslinked gel has macropores of average size of about 700nm. In certain embodiments, the macroporous crosslinked gel hasmacropores of average size of about 300 nm.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein themacroporous cross-linked gel of the composite material is a hydrogel, apolyelectrolyte gel, a hydrophobic gel, a neutral gel, or a gelcomprising functional groups. In certain embodiments, the inventionrelates to any one of the above-mentioned fluid treatment devices orelements, wherein the macroporous cross-linked gel of the compositematerial is a neutral or charged hydrogel; and the neutral or chargedhydrogel is selected from the group consisting of cross-linkedpoly(vinyl alcohol), poly(acrylamide), poly(isopropylacrylamide),poly(vinylpyrrolidone), poly(hydroxymethyl acrylate), poly(ethyleneoxide), copolymers of acrylic acid or methacrylic acid with acrylamide,isopropylacrylamide, or vinylpyrrolidone, copolymers ofacrylamide-2-methyl-1-propanesulfonic acid with acrylamide,isopropylacrylamide, or vinylpyrrolidone, copolymers of(3-acrylamido-propyl) trimethylammonium chloride with acrylamide,isopropylacrylamide, or N-vinyl-pyrrolidone, and copolymers ofdiallyldimethylammonium chloride with acrylamide, isopropylacrylamide,or vinylpyrrolidone. In certain embodiments, the invention relates toany one of the above-mentioned fluid treatment devices or elements,wherein the macroporous cross-linked gel of the composite material is apolyelectrolyte gel; and the polyelectrolyte gel is selected from thegroup consisting of cross-linkedpoly(acrylamido-2-methyl-1-propanesulfonic acid) and its salts,poly(acrylic acid) and its salts, poly(methacrylic acid) and its salts,poly(styrenesulfonic acid) and its salts, poly(vinylsulfonic acid) andits salts, poly(alginic acid) and its salts,poly[(3-acrylamidopropyl)trimethylammonium] salts,poly(diallyldimethylammonium) salts, poly(4-vinyl-N-methylpyridinium)salts, poly(vinylbenzyl-N-trimethylammonium) salts, andpoly(ethyleneimine) and its salts. In certain embodiments, the inventionrelates to any one of the above-mentioned fluid treatment devices orelements, wherein the macroporous cross-linked gel of the compositematerial is a hydrophobic gel; and the hydrophobic gel is selected fromthe group consisting of cross-linked polymers or copolymers of ethylacrylate, n-butyl acrylate, propyl acrylate, octyl acrylate, dodecylacrylate, octadecylacrylamide, stearyl acrylate, and styrene. In certainembodiments, the invention relates to any one of the above-mentionedfluid treatment devices or elements, wherein the macroporouscross-linked gel of the composite material is a neutral gel; and theneutral gel is selected from the group consisting of cross-linkedpolymers or copolymers of acrylamide, N,N-dimethylacrylamide,N-methacryloylacrylamide, N-methyl-N-vinylacetamide, andN-vinylpyrrolidone.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein themacroporous cross-linked gel of the composite material is a gelcomprising functional groups. In certain embodiments, the macroporouscross-linked gel of the composite material comprises monomers, whereinthe monomers comprise functional groups. In certain embodiments, thefunctional groups are thiols or protected thiols. In certainembodiments, the macroporous cross-linked gel comprises monomers,wherein the monomers are selected from the group consisting of allyl3-mercaptopropionate thioacetate,(S-benzoyl-3-mercapto-2-hydroxypropyl)-2-methyl-2-propenoate,(S-2,2-dimethylpropanoyl-3-mercapto-2-hydroxypropyl)-2-methyl-2-propenoate,(S-acetyl-3-mercapto-2-acetylpropyl)-2-methyl-2-propenoate,(S-acetyl-3-mercapto-2-hydroxypropyl)-2-methyl-2-propenoate,(S-acetyl-3-mercapto-2-acetoacetylpropyl)-2-methyl-2-propenoate,(S-acetyl-3-mercapto-2-tetrahydropyranyl)-2-methyl-2-propenoate,(S-acetyl-3-mercapto-2-(2-methoxy-2-propoxy))-2-methyl-2-propenoate,(S-acetyl-2-mercapto-3-acetylpropyl)-2-methyl-2-propenoate,S-acetyl-(1-allyloxy-3-mercapto-2-hydroxypropane),S-benzoyl-(1-allyloxy-3-mercapto-2-hydroxypropane) andS-2,2-dimethylpropanoyl-(1-allyloxy-3-mercapto-2-hydroxypropane).

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises functional groups; and the functionalgroups are selected from the group consisting of amino acid ligands,antigen and antibody ligands, dye ligands, biological molecules,biological ions, and metal affinity ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises functional groups; and said functionalgroups are metal affinity ligands. In certain embodiments, the inventionrelates to any one of the above-mentioned fluid treatment devices orelements, wherein the composite material comprises functional groups;said functional groups are metal affinity ligands; and a plurality ofmetal ions are complexed to a plurality of said metal affinity ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are polydentate ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are octadentate, hexadentate, tetradentate, tridentateor bidentate ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are tetradentate ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are tridentate ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are bidentate ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are iminodicarboxylic acid ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are iminodiacetic acid ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are salts of iminodiacetic acid ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are sodium salts of iminodiacetic acid ligands. See,e.g., FIG. 24.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands are potassium salts of iminodiacetic acid ligands.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands comprise ethylenediamine moieties. See, e.g., FIG. 25.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands comprise hexamethylenediamine moieties. See, e.g., FIG.26.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands comprise diethanolamine moieties. See, e.g., FIG. 27.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands comprise pentaethylenehexamine moieties. See, e.g.,FIG. 28.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands comprise triethylenetetramine moieties. See, e.g., FIG.29.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands comprise tris(carboxymethyl)ethylene diamine. See,e.g., FIG. 30.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands; and said metalaffinity ligands comprise conjugate bases of carboxylic acids. Incertain embodiments, the conjugate bases are available as salts. Incertain embodiments, the conjugate bases are available as sodium saltsor potassium salts. In certain embodiments, the conjugate bases areavailable as sodium salts. In certain embodiments, the conjugate basesare available as potassium salts.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; and said metal ions are transition metal ions,lanthanide ions, poor metal ions or alkaline earth metal ions.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; and said metal ions are selected from the groupconsisting of nickel, zirconium, lanthanum, cerium, manganese, titanium,cobalt, iron, copper, zinc, silver, gallium, platinum, palladium, lead,mercury, cadmium and gold.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; and said metal ions are nickel or zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; and said metal ions are nickel.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; and said metal ions are zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are octadentate,hexadentate, tetradentate, tridentate or bidentate ligands; and saidmetal ions are transition metal ions, lanthanide ions, poor metal ionsor alkaline earth metal ions.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are octadentate,hexadentate, tetradentate, tridentate or bidentate ligands; and saidmetal ions are selected from the group consisting of nickel, zirconium,lanthanum, cerium, manganese, titanium, cobalt, iron, copper, zinc,silver, gallium, platinum, palladium, lead, mercury, cadmium and gold.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are octadentate,hexadentate, tetradentate, tridentate or bidentate ligands; and saidmetal ions are nickel or zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are octadentate,hexadentate, tetradentate, tridentate or bidentate ligands; and saidmetal ions are nickel.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are octadentate,hexadentate, tetradentate, tridentate or bidentate ligands; and saidmetal ions are zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tetradentateligands; and said metal ions are transition metal ions, lanthanide ions,poor metal ions or alkaline earth metal ions.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tetradentateligands; and said metal ions are selected from the group consisting ofnickel, zirconium, lanthanum, cerium, manganese, titanium, cobalt, iron,copper, zinc, silver, gallium, platinum, palladium, lead, mercury,cadmium and gold.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tetradentateligands; and said metal ions are nickel or zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tetradentateligands; and said metal ions are nickel.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tetradentateligands; and said metal ions are zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tridentateligands; and said metal ions are transition metal ions, lanthanide ions,poor metal ions or alkaline earth metal ions.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tridentateligands; and said metal ions are selected from the group consisting ofnickel, zirconium, lanthanum, cerium, manganese, titanium, cobalt, iron,copper, zinc, silver, gallium, platinum, palladium, lead, mercury,cadmium and gold.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tridentateligands; and said metal ions are nickel or zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tridentateligands; and said metal ions are nickel.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are tridentateligands; and said metal ions are zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are bidentateligands; and said metal ions are transition metal ions, lanthanide ions,poor metal ions or alkaline earth metal ions.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are bidentateligands; and said metal ions are selected from the group consisting ofnickel, zirconium, lanthanum, cerium, manganese, titanium, cobalt, iron,copper, zinc, silver, gallium, platinum, palladium, lead, mercury,cadmium and gold.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are bidentateligands; and said metal ions are nickel or zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are bidentateligands; and said metal ions are nickel.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are bidentateligands; and said metal ions are zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands areiminodicarboxylic acid ligands; and said metal ions are transition metalions, lanthanide ions, poor metal ions or alkaline earth metal ions.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands areiminodicarboxylic acid ligands; and said metal ions are selected fromthe group consisting of nickel, zirconium, lanthanum, cerium, manganese,titanium, cobalt, iron, copper, zinc, silver, gallium, platinum,palladium, lead, mercury, cadmium and gold.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands areiminodicarboxylic acid ligands; and said metal ions are nickel orzirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands areiminodicarboxylic acid ligands; and said metal ions are nickel.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands areiminodicarboxylic acid ligands; and said metal ions are zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are iminodiaceticacid ligands; and said metal ions are transition metal ions, lanthanideions, poor metal ions or alkaline earth metal ions.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are iminodiaceticacid ligands; and said metal ions are selected from the group consistingof nickel, zirconium, lanthanum, cerium, manganese, titanium, cobalt,iron, copper, zinc, silver, gallium, platinum, palladium, lead, mercury,cadmium and gold.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are iminodiaceticacid ligands; and said metal ions are nickel or zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are iminodiaceticacid ligands; and said metal ions are nickel.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises metal affinity ligands complexed to aplurality of metal ions; said metal affinity ligands are iminodiaceticacid ligands; and said metal ions are zirconium.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises functional groups; and the functionalgroups are biological molecules or biological ions. In certainembodiments, the biological molecules or biological ions are selectedfrom the group consisting of albumins, lysozyme, viruses, cells,γ-globulins of human and animal origins, immunoglobulins of both humanand animal origins, proteins of recombinant or natural origin including,polypeptides of synthetic or natural origin, interleukin-2 and itsreceptor, enzymes, monoclonal antibodies, antigens, lectins, bacterialimmunoglobulin-binding proteins, trypsin and its inhibitor, cytochromeC, myoglobulin, recombinant human interleukin, recombinant fusionprotein, Protein A, Protein G, Protein L, Peptide H, nucleic acidderived products, DNA of either synthetic or natural origin, and RNA ofeither synthetic or natural origin.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material comprises Protein A. Protein A is a 40-60 kDa MSCRAMMsurface protein originally found in the cell wall of the bacteriaStaphylococcus aureus. It is encoded by the spa gene and its regulationis controlled by DNA topology, cellular osmolarity, and a two-componentsystem called ArlS-ArlR. It has found use in biochemical researchbecause of its ability to bind immunoglobulins. It binds proteins frommany of mammalian species, most notably IgGs. It binds with the Fcregion of immunoglobulins through interaction with the heavy chain. Theresult of this type of interaction is that, in serum, the bacteria willbind IgG molecules in the wrong orientation (in relation to normalantibody function) on their surface which disrupts opsonization andphagocytosis. It binds with high affinity to human IgG1 and IgG2 as wellas mouse IgG2a and IgG2b. Protein A binds with moderate affinity tohuman IgM, IgA and IgE as well as to mouse IgG3 and IgG1. It does notreact with human IgG3 or IgD, nor will it react to mouse IgM, IgA orIgE.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein themacroporous crosslinked gel of the composite material comprises amacromonomer. In certain embodiments, the macromonomer is selected fromthe group consisting of poly(ethylene glycol) acrylate and poly(ethyleneglycol) methacrylate.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein themacroporous cross-linked gel of the composite material is cross-linkedby N,N-methylenebisacrylamide or a polyfunctional macromonomer. Incertain embodiments, the macroporous cross-linked gel of the compositematerial is cross-linked by a polyfunctional macromonomer; and thepolyfunctional macromonomer is selected from the group consisting ofpoly(ethylene glycol) diacrylate and poly(ethylene glycol)dimethacrylate. In certain embodiments, the invention relates to any oneof the above-mentioned fluid treatment devices or elements, wherein themacroporous cross-linked gel of the composite material is cross-linkedby N,N-methylenebisacrylamide.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein themacroporous cross-linked gel of the composite material is a positivelycharged hydrogel comprising a co-polymer of(3-acrylamidopropyl)trimethylammonium chloride (APTAC) andN-(hydroxymethyl)acrylamide cross-linked by N,N′-methylenebisacrylamide.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material is a membrane; and the macroporous cross-linked gelbears charged moieties.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite material is a membrane for use as a filter in size exclusionseparation.

In certain embodiments, the fluid treatment devices or elements of theinvention comprise any one of the above-mentioned composite materials,wherein the composite materials comprise negatively-charged moieties.Negatively-charged membranes repel foulants at the membrane surfaceresulting in higher flux, easier cleanings, and lower system costs.

In certain embodiments, the fluid treatment devices or elements of theinvention comprise any one of the above-mentioned composite materials,wherein the composite materials are hydrophilic in nature. Foulants aretypically hydrophobic species.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein the supportmember of the composite material consists essentially of polymericmaterial in the form of a membrane that has a thickness of from about 10μm to about 500 μm and comprises pores of average size between about 0.1to about 25 μm.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein the supportmember of the composite material consists essentially of a polyolefin.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein the supportmember of the composite material comprises a polymeric material selectedfrom the group consisting of polysulfones, polyethersulfones,polyphenyleneoxides, polycarbonates, polyesters, cellulose and cellulosederivatives.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein the supportmember of the composite material consists essentially of polymericmaterial in the form of a fibrous fabric that has a thickness of fromabout 10 μm to about 2000 μm and comprises pores of average size fromabout 0.1 to about 25 μm.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein the supportmember of the composite material comprises a stack of 2 to 10 separatesupport members.

In certain embodiments, the invention relates to any one of theabove-mentioned fluid treatment devices or elements, wherein thecomposite materials are disk-shaped, thereby forming cut disk membranes.In certain embodiments, the cut disk membranes are from about 5 mm indiameter to about 100 mm in diameter. In certain embodiments, the cutdisk membranes are from about 10 mm in diameter to about 75 mm indiameter. In certain embodiments, the cut disk membranes are from about15 mm in diameter to about 55 mm in diameter. In certain embodiments,the cut disk membranes are about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 mm in diameter. Incertain embodiments, the cut disk membranes are about 18 mm in diameter.In certain embodiments, the cut disk membranes are about 25 mm indiameter. In certain embodiments, the cut disk membranes are about 50 mmin diameter. In certain embodiments, the cut disk membranes are made bysimply cutting from sheets of composite material.

Exemplary Methods

In certain embodiments, the invention relates to a method comprising thestep of:

-   -   contacting a first fluid comprising a substance with a composite        material in any one of the above-mentioned fluid treatment        devices, thereby adsorbing or absorbing the substance onto the        composite material.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, further comprising the step of

-   -   placing the first fluid in an inlet of the fluid treatment        device.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the first fluid is passed along a fluidflow path substantially perpendicular to the pores of the supportmember.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, further comprising the step of

-   -   contacting a second fluid with the substance adsorbed or        absorbed onto the composite material, thereby releasing the        substance from the composite material.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the second fluid is passed through themacropores of the composite material, thereby releasing the substancefrom the composite material.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the second fluid is passed along thefluid flow path substantially perpendicular to the pores of the supportmember, thereby releasing the substance from the composite material.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the substance is separated based onsize exclusion.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the macroporous gel displays a specificinteraction for the substance.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the specific interactions areelectrostatic interactions, affinity interactions, or hydrophobicinteractions.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the specific interactions areelectrostatic interactions, the composite material bears charges on themacroporous gel; the substance is charged; and the substance isseparated based on Donnan exclusion.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the first fluid is a suspension ofcells or a suspension of aggregates.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the substance is a biological moleculeor biological ion.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the biological molecule or biologicalion is selected from the group consisting of albumins, lysozyme,viruses, cells, γ-globulins of human and animal origins, immunoglobulinsof both human and animal origins, proteins of recombinant or naturalorigin including, polypeptides of synthetic or natural origin,interleukin-2 and its receptor, enzymes, monoclonal antibodies, trypsinand its inhibitor, cytochrome C, myoglobulin, recombinant humaninterleukin, recombinant fusion protein, nucleic acid derived products,DNA of either synthetic or natural origin, and RNA of either syntheticor natural origin.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the biological molecule or biologicalion is a protein; and the protein comprises exposed amino acid residuesselected from the group consisting of Glu, Asp, Try, Arg, Lys, Met, andHis.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the biological molecule or biologicalion is a protein; and the protein comprises exposed His amino acidresidues.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the biological molecule or biologicalion is a monoclonal antibody.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the substance is a metal-containingparticle, or a metal-containing ion.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the metal-containing particle ormetal-containing ion comprises a transition metal, a lanthanide, a poormetal, or an alkaline earth metal.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the metal-containing particle ormetal-containing ion comprises a metal selected from the groupconsisting of nickel, zirconium, lanthanum, cerium, manganese, titanium,cobalt, iron, copper, zinc, silver, gallium, platinum, palladium, lead,mercury, cadmium and gold.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the first fluid is waste water.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the first fluid is waste water from orerefining, or seawater.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the substance is lead or mercury.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the substance is platinum, palladium,copper, gold, or silver.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the fluid is waste water; and themetal-containing particle or metal-containing ion comprises lead ormercury.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the first fluid is waste water from orerefining; and the metal-containing particle or metal-containing ioncomprises lead or mercury.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the first fluid is seawater; and themetal-containing particle or metal-containing ion comprises platinum,palladium, copper, gold, or silver.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the first fluid comprises egg white.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the first fluid comprises egg white;and the substance is lysozyme.

In certain embodiments, the invention relates to a method in which, intangential flow separation mode, no pre-processing of the raw reactionmixtures is required due to the high specificity of the compositematerials in the devices of the present invention. In certainembodiments, the invention relates to a method in which separations canbe carried out on a large scale. In certain embodiments, the inventionrelates to a method in which separations can be carried out in a shorteramount of time. In certain embodiments, the invention relates to amethod in which the devices have a high binding capacity.

In certain embodiments, the invention relates to a method that comprisestwo steps—collecting the desired substance onto the composite materialand harvesting the desired substance from the composite material. Incertain embodiments, the first step is run in tangential separationmode. In certain embodiments, the first step is run in tangentialseparation mode and the second step is run in direct filtration modewith a second fluid.

In certain embodiments, the invention relates to a method of separatinga substance from a fluid, comprising the step of:

placing the fluid in contact with a composite material in any one of theabove-mentioned fluid treatment devices, thereby adsorbing or absorbingthe substance to the composite material.

In certain embodiments, the invention relates to a method of separatinga substance from a fluid, comprising the step of:

placing the fluid in an inlet of any one of the above-mentioned fluidtreatment devices, thereby adsorbing or absorbing the substance to thecomposite material and producing a permeate; and

collecting the permeate from an outlet of the fluid treatment device.

In certain embodiments, the invention relates to the above-mentionedmethod, wherein the fluid is passed over the surface of the compositematerial; and the substance is adsorbed or absorbed onto the surface ofthe composite material.

In certain embodiments, the invention relates to the above-mentionedmethod, wherein the fluid is passed through the macropores of thecomposite material; and the substance is adsorbed or absorbed within themacropores of the composite material.

In certain embodiments, the invention relates to a method of separatinga substance from a fluid, comprising the step of:

placing the fluid in an inlet of any one of the above-mentioned fluidtreatment devices, thereby adsorbing or absorbing the substance to thecomposite material;

collecting the permeate from an outlet of the fluid treatment device;

placing a second fluid in the inlet of the fluid treatment device,thereby releasing the substance from the composite material.

In certain embodiments, the invention relates to the above-mentionedmethod, wherein the fluid is passed over the surface of the compositematerial; the substance is adsorbed or absorbed onto the surface of thecomposite material; and the second fluid is passed through themacropores of the composite material, thereby releasing the substancefrom the composite material.

In certain embodiments, the invention relates to the above-mentionedmethod, wherein the fluid is passed over the surface of the compositematerial; the substance is adsorbed or absorbed onto the surface of thecomposite material; and the second fluid is passed over the surface ofthe composite material, thereby releasing the substance from the surfaceof the composite material.

In certain embodiments, the invention relates to the above-mentionedmethod, wherein the fluid is passed through the macropores of thecomposite material; the substance is adsorbed or absorbed within themacropores of the composite material; and the second fluid is passedover the surface of the composite material, thereby releasing thesubstance from the composite material.

In certain embodiments, the invention relates to the above-mentionedmethod, wherein the fluid is passed through the macropores of thecomposite material; the substance is adsorbed or absorbed within themacropores of the composite material; and the second fluid is passedthrough the macropores of the composite material, thereby releasing thesubstance from the composite material.

In certain embodiments, the invention relates to any one of theabove-mentioned methods, wherein the substance is radioactive.

In certain embodiments, the invention relates to a method of separatinga substance from a fluid, comprising the steps of:

placing the fluid in an inlet of any one of the above-mentioned fluidtreatment devices, thereby adsorbing or absorbing the substance to thecomposite material and producing a permeate; and

collecting the permeate from an outlet of the fluid treatment device,

wherein the fluid comprises egg white; and the substance is lysozyme.

In certain embodiments, the invention relates to the above-mentionedmethod, wherein the fluid is passed over the surface of the fluidtreatment element; and the substance is adsorbed or absorbed onto thesurface of the fluid treatment element.

In certain embodiments, the invention relates to a method of separatinga substance from a first fluid, comprising the steps of:

placing the first fluid in an inlet of any one of the above-mentionedfluid treatment devices, thereby adsorbing or absorbing the substance tothe composite material;

collecting the permeate from an outlet of the fluid treatment device;

placing a second fluid in the inlet of the fluid treatment device,thereby releasing the substance from the composite material,

wherein the first fluid comprises egg white; and the substance islysozyme.

In certain embodiments, the invention relates to the above-mentionedmethod, wherein the fluid is passed over the surface of the fluidtreatment element; the substance is adsorbed or absorbed onto thesurface of the fluid treatment element; and the second fluid is passedthrough the macropores of the fluid treatment element, thereby releasingthe substance from the macropores. In certain embodiments, the inventionrelates to the above-mentioned method, wherein the fluid is passed overthe surface of the fluid treatment element; the substance is adsorbed orabsorbed onto the surface of the fluid treatment element; and the secondfluid is passed over the surface of the fluid treatment element, therebyreleasing the substance from the surface of the fluid treatment element.

EXEMPLIFICATION

The invention now being generally described, it will be more readilyunderstood by reference to the following examples, which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention, and are not intended to limit the invention.

Example 1

Dead-End Versus Cross-Flow Modes for Viral Capture

An example of the improvement of cross-flow technology versus dead-endtechnology can be realized when the two modes are compared directly toeach other. FIG. 1 shows two experiments in which a specific device wasrun as a dead-end and as cross-flow device. The material of interest isa virus. In both cases, the capture of the cross-flow device exceededthe dead-end version, as indicated by the amount to the pure targetmaterial capture after washing and elution.

Example 2

Chromatographic Capture and Harvest: Elution of Ovalbumin and Lysozyme

The membrane can selectively adsorb two protein materials from the feedstream and then, through the use of an altering buffer fluid,selectively elute the target bio-molecules. FIGS. 2 and 3 illustratethis effect. The initial feed stream of egg white was exposed to amembrane surface in cross-flow mode. Once the feed stream was removedand the membrane washed, both Ovalbumin and lysozyme were found adheredto the membrane (FIG. 2). Under specific buffer conditions, the proteinswere selectively eluted (FIG. 3), which demonstrates the chromatographicnature of the membrane in cross-flow mode.

Example 3

Orthogonal Two-Step Capture and Harvest

See FIG. 4.

Example 4

Device Design: Wrap Design Data and Schematics

The effect of selecting an optimal spacer material in a simple wrappeddesign was observed and is depicted in FIG. 5. In this design, a rollwas made by layering the necessary membrane sheet between two identicalspacer sheets and rolling the multi-layered structure into a column.This column was then placed into a metal tube housing that was fittedwith end-caps which have both an inlet(s) and outlet(s) attached (FIG.6). The larger spacer material and loose-wind structure enabled theideal cross-flow or tangential-flow adsorption. Importantly, this designeliminated any direct trans-membrane flow, as the process fluid was runon either side of the membrane. Thus, the improvements stemmed, at leastpartially, from the low shear environment. The purified lysozyme controlwas run on the device and was used to represent 100% or maximumadsorption of the target species. The remaining data were generateddirectly from process fluid streams (egg whites). In this embodiment,the spacer layer materials on either side of the membrane were identicalbut there is no requirement for this symmetry and the “roll” could havediffering layers or one layer could be completely absent.

Example 5

Device Design: Cassettes

The height of the feed channel may impact the ability to maximize theamount of adsorbed target molecule. Smaller feed channel heights mayinduce greater shear or turbulence at the membrane surface, which eitherinhibits adsorption or removes target material that does deposit. Thechannel height needs to be at least 10 mm and more ideally >20 mm,typically 23 mm (FIG. 7).

Screen changes have not been identified as a driver for performance incassette design.

Example 6

Device Design: Spiral

FIG. 23 depicts a spiral wound device. When membranes of the currentinvention are incorporated into this device, highly contaminated or veryviscous feed streams can be effectively separated into their desiredparts.

Example 7

Antibody Purification: Membrane Functionalized with Protein A

A 0.01 SQM Protein A cassette with an open channel, suspended screendesign which enabled a fluid flow tangential to the plane of themembrane was evaluated using an un-clarified feed stream which containedthe monoclonal antibody (mAb) target. Traditional resin chromatographicseparation processes cannot process un-clarified feed streams. The onlymethod that had been demonstrated able to capture the mAb target onbench scale was an expanded bed column functioning in batch mode with astatic soak. This modified expanded bed was uneconomical and impracticalat larger scale. On the other hand, the cassettes were effective incapturing the target MAb product when used a simple flow through mode.This mode allowed the process stream to flow across the membrane suchthat debris in the fluid did not blind the membrane. Binding of thetarget species in this mode was a surface effect only.

Lysis Procedure:

270 g mAb 4420 pellet diluted 1 part pellet, 3 parts 10× phosphatebuffer solution (PBS), 1 part 5× Pfenix lysis buffer. Homogenized for 5minutes, and sonicated for 10 minutes. Loaded onto the membrane as anun-clarified and undiluted feed stream.

Membrane Procedure:

A membrane cassette with an active surface area of 0.01 m² and pore sizeof 0.3 μm was equilibrated in PBS at pH 7.4. This device was then loadedby complete system recirculation with lysed mAb 4420 for one hour. Thedevice was then washed with 1 L of 1×PBS pH 7.4, and eluted using a 10minute recirculation of 0.1 M Glycine at pH 2.9 followed by 100 mLsystem flush with 0.1 M Glycine pH 2.9. The feed was run in flow throughmode at 100 mL/min with permeate shut off which limited the device tosurface binding only from the un-clarified lysate. A gel electrophoresisqualitative analysis indicated that significant amounts of mAb had beencaptured.

Conclusion:

The cross-flow product provided a simple, on-off bind-elute captureprocess that could capture and concentrate intact mAb (observed bindingwas in the range of 5-10 mg/mL) as well as a lot of contaminants. Withdevelopment, membrane could serve as a scalable capture method forun-clarified mAb Pseudomonas feed streams.

Example 8

His-Tagged Protein Purification: Membrane Functionalized with IMAC Ni

A 0.02 SQM IMAC-Ni (iminodiacetic acid complexed to Ni) cassette with anopen channel, suspended screen design which enabled a fluid flowtangential to the plane of the membrane was evaluated using a feedstream containing a his-tagged protein target. Traditional resin-basedchromatographic separation processes cannot process un-clarified feedstreams. In this experiment, the cassettes were effective in capturingthe target product in a simple flow through mode. This mode allowed theprocess stream to flow across the membrane such that debris in the fluiddid not blind the membrane. Binding of the target species in this modewas a surface effect only. The product was able to be run with multiplecycles with no loss in binding capacity.

Lysis Procedure:

8 L of cell harvest material was diluted with 2 L of 5× Pfenix LysisBuffer. This mixture was allowed to mix for 2 hours until the materialhad liquefied.

Membrane Procedure:

A membrane cassette with an active surface area of 0.02 m² wasequilibrated in 50 mM PBS, 500 mM NaCl, 5% (wt) glycerol and 25 mMImidazole at pH 8.0. The cassette was then run with the liquefied cellharvest material at a 100 mL/min feed rate with no permeate flow. Thiswas flowed by elution using 500 mL 1×PBS, 500 mM Imidazole at pH 7.4.This process was repeated 3 times in sequence with the follow details:

-   -   Run 1: 1 L of Lysate was centrifuged and diluted by a factor of        5 in equilibration buffer. The device was then loaded by        complete system recirculation for 1 hour time and material was        able to permeate through the pores at 20 PSIg inlet pressure. A        gel electrophoresis qualitative analysis indicated that        significant amounts his-tagged protein had been captured.    -   Run 2: 1 L of Lysate was only centrifuged prior to use. The        device was then loaded by complete system recirculation for 1        hour time and material was able to permeate through the pores at        20 PSIg inlet pressure. A gel electrophoresis qualitative        analysis indicated that significant amounts his-tagged protein        had been captured    -   Run 3: 1 L of Lysate was used without pre-treatment. The device        was then loaded by complete system recirculation for 15, 30, 45,        60 minute load times. The device was eluted after each cycle and        the membrane not stripped (cleaned) between elutions. When the        permeate line was closed, no back pressure increase to system        was observed during load indicating that the membrane had not        blinded.

Conclusion:

The IMAC-Ni product yielded excellent purification characteristics(observed binding was in the range of 60-80 mg/mL) without optimization.Despite manufacturers designation as “single-use,” 7× reuse demonstratedfor IMAC membrane without the need for EDTA strip and re-charge.

INCORPORATION BY REFERENCE

All of the U.S. patents and U.S. patent application publications citedherein are hereby incorporated by reference.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

We claim:
 1. A method, comprising the steps of: 1) contacting a firstfluid, comprising a substance, with a composite material; wherein: thecomposite material comprises: a support member, comprising a pluralityof pores extending through the support member; and a cross-linked gel;the first fluid is an unclarified feed stream; the substance is amonoclonal antibody; the cross-linked gel is selected from the groupconsisting of N,N′-methylenebisacrylamide cross-linked copolymers orcross-linked polymers of isopropylacrylamide, dodecyl acrylate, ethylacrylate, ethylene oxide, hydroxymethyl acrylate, n-butyl acrylate,N-(hydroxymethyl)acrylamide, N-methacryloylacrylamide,N-methyl-N-vinylacetamide, N,N-dimethylacrylamide, N-vinyl-pyrrolidone,octadecylacrylamide, octyl acrylate, propyl acrylate, stearyl acrylate,styrene, and vinyl alcohol, or a mixture thereof; the cross-linked gelcomprises functional groups, wherein said functional groups are ProteinA; the cross-linked gel is located in the pores of the support member;and the pores of the support member are substantially perpendicular tothe fluid flow path between an inlet and a retentate outlet, and saidfluid flow path between the inlet and the retentate outlet issubstantially parallel to the surface of a filter medium comprising thecomposite material; thereby adsorbing or absorbing the monoclonalantibody onto the composite material; and 2) contacting a second fluidwith the monoclonal antibody adsorbed or absorbed onto the compositematerial, thereby releasing the monoclonal antibody from the compositematerial.
 2. The method of claim 1, wherein the first fluid is asuspension of cells or a suspension of aggregates.
 3. The method ofclaim 1, wherein the first fluid is more viscous than water.
 4. Themethod of claim 1, wherein the first fluid is an unclarified Pseudomonasfeed stream.
 5. The method of claim 1, wherein the cross-linked gel isselected from the group consisting of N,N′-methylenebisacrylamidecross-linked copolymers or cross-linked polymers of isopropylacrylamide,N-(hydroxymethyl)acrylamide, N-methacryloylacrylamide,N-methyl-N-vinylacetamide, N,N-dimethylacrylamide, N-vinyl-pyrrolidone,octadecylacrylamide, styrene, and vinyl alcohol, or a mixture thereof.6. The method of claim 1, wherein the cross-linked gel is selected fromthe group consisting of N,N′-methylenebisacrylamide cross-linkedcopolymers or cross-linked polymers of vinyl alcohol,isopropylacrylamide, N-vinylpyrrolidone, hydroxymethyl acrylate,ethylene oxide, copolymers of acrylic acid or methacrylic acid withisopropylacrylamide, or N-vinylpyrrolidone, copolymers ofacrylamide-2-methyl-1-propanesulfonic acid with isopropylacrylamide orN-vinylpyrrolidone, copolymers of (3-acrylamido-propyl)trimethylammonium chloride with isopropylacrylamide orN-vinyl-pyrrolidone, and copolymers of diallyldimethylammonium chloridewith isopropylacrylamide or N-vinylpyrrolidone.
 7. The method of claim1, wherein the cross-linked gel is selected from the group consisting ofN,N′-methylenebisacrylamide cross-linked copolymers or cross-linkedpolymers of ethyl acrylate, n-butyl acrylate, propyl acrylate, octylacrylate, dodecyl acrylate, octadecylacrylamide, stearyl acrylate, andstyrene.
 8. The method of claim 1, wherein the cross-linked gel isselected from the group consisting of N,N′-methylenebisacrylamidecross-linked copolymers or cross-linked polymers ofN,N-dimethylacrylamide, N-methacryloylacrylamide,N-methyl-N-vinylacetamide, and N-vinylpyrrolidone.
 9. The method ofclaim 1, wherein the cross-linked gel is macroporous.
 10. The method ofclaim 1, wherein the composite material is a membrane.